Microbial Control of Host Intercellular Communication

宿主细胞间通讯的微生物控制

基本信息

批准号：
10661500
负责人：
REBECCA L LAMASON
金额：
$ 30.56万
依托单位：
MASSACHUSETTS INSTITUTE OF TECHNOLOGY
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-07-15 至 2026-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10661500
关键词：
Acceleration Actins Adhesions Bacteria Binding Biological Biology Buffers CAV1 gene CAV2 gene Cardiovascular Diseases Caveolae Caveolins Cell Communication Cell physiology Cell-Cell Adhesion Cells Cellular biology Communication Communications Media Complex Cytoskeleton Development Disease Endocytosis Eukaryotic Cell Genes Goals Health Homeostasis Human Image Immunology Intercellular Junctions Life Listeria Listeria monocytogenes Malignant Neoplasms Mechanical Stress Mechanics Mediating Membrane Microbe Microscopy Molecular Molecular Probes Organelles Organism Pathway interactions Polymers Process Protein Dynamics Proteins Quantitative Microscopy RNA Interference RNA interference screen Regulation Role Shapes Signal Transduction Small Interfering RNA Structure Tail Testing Tissues Work cell fixing cell motility host-microbe interactions human disease human tissue improved innovation insight intercellular communication knock-down mechanical signal mechanotransduction microbial novel pathogenic bacteria recruit response spatiotemporal tool

项目摘要

PROJECT SUMMARY/ABSTRACT Multicellular organisms use intercellular communication to coordinate cell function and maintain tissue homeostasis. Recent work suggests that this communication is driven in part by the exchange of organelles (via trans-endocytosis) and mechanical cues directly across cell-cell junctions. Dysregulation of this communication leads to cancer and cardiovascular diseases. Despite its importance, we lack a fundamental molecular understanding of how intercellular communication occurs because of the limited number of cell biological tools capable of probing the molecular mechanisms at cell-cell contacts. This proposal seeks to elucidate the regulatory mechanisms of the pathways thought to control intercellular communication by studying how they are manipulated when under microbial control. The bacterium Listeria monocytogenes disseminates through human tissues using a process called cell-to-cell spread, which is a vesicular-mediated form of intercellular exchange that mimics host trans-endocytosis. Listeria spreads from cell to cell by mobilizing the host’s actin cytoskeleton for intracellular motility and transport to the cell-cell junction. Once at the junction, it pushes against the membrane and forms a double-membrane protrusion that is engulfed by a neighboring cell. Studying this distinctive spreading process will allow us to examine several outstanding cell biological questions. First, are specific endocytic pathways used at cell-cell junctions to engulf large cargo like microbes? Second, are mechanically-sensitive membrane domains or membrane curvature proteins activated as Listeria pushes against the junction during spread? To answer these questions, we used a high-content, image-based siRNA screen to test if Listeria requires host intercellular communication pathways during spread. We discovered that the endocytic and mechanoresponsive caveolar proteins CAV1, CAV2, and PACSIN2 promote Listeria spread. We also revealed a putative role for 19 other host proteins, including those that regulate membrane curvature, trans- endocytosis, and adhesion. Our preliminary findings suggest the overall hypothesis that Listeria subverts multiple intercellular communication pathways to promote cell-to-cell spread. In Aim 1, we will determine how PACSIN2 and caveolins coordinate their activities to promote the engulfment stage of cell-to-cell spread. In Aim 2, we will reveal which of the remaining hits regulate Listeria spread specifically, how they function, and if they work independently or together with caveolae. In the end, our proposed studies will improve our fundamental understanding of host-microbe interactions and basic cell biology, and may uncover how intercellular communication goes awry in human disease.

项目摘要/摘要多细胞生物使用细胞间通信来协调细胞功能并维持组织稳态。最近的工作表明，这种交流部分是由细胞器交换所驱动的（通过直接跨细胞 - 细胞连接的跨性腺细胞增多症）和机械提示。这种交流的失调导致癌症和心血管疾病。尽管它很重要，但我们缺乏基本分子由于细胞生物学工具数量有限，了解细胞间通信的发生方式能够探测细胞细胞接触处的分子机制。该建议旨在阐明被认为是通过研究如何控制细胞间通信的途径的调节机制在微生物控制下进行操纵。李斯特菌单核细胞增生李斯特菌通过人传播使用称为细胞到细胞扩散的过程，这是囊泡介导的细胞间交换形式该模仿宿主的跨性腺细胞增多症。李斯特菌通过动员宿主的肌动蛋白细胞骨架从细胞传播到细胞用于细胞内运动和运输到细胞 - 细胞连接。一旦在交界处，它就会推向膜并形成一个被相邻细胞吞没的双膜突出。研究这个独特的扩散过程将使我们能够检查几个出色的细胞生物学问题。首先，是在细胞 - 细胞连接处使用的特定内吞途径吞噬了大型货物，例如微生物？第二，是机械敏感的膜结构域或膜曲率蛋白被激活，因为李斯特菌推向传播期间的交界处？为了回答这些问题，我们使用了一个基于图像的siRNA屏幕测试李斯特菌是否需要在扩散过程中宿主的细胞间通信。我们发现内吞和机械辅助洞穴蛋白Cav1，Cav2和Pacsin2促进李斯特菌扩散。我们还揭示了其他19种宿主蛋白的推定作用，包括调节膜曲率的蛋白质内吞作用和粘膜。我们的初步发现表明了总体假设，即李斯特菌颠覆了多个促进细胞间扩散的细胞间通信途径。在AIM 1中，我们将确定PACSIN 2 小窝蛋白会协调其活动，以促进细胞对细胞扩散的吞噬阶段。在AIM 2中，我们将揭示其余哪些命中率调节李斯特菌的特殊传播，它们的运作方式以及是否起作用独立或与小屋一起。最后，我们拟议的研究将改善我们的基本了解宿主 - 微生物相互作用和基本细胞生物学，并可能发现细胞间人类疾病的沟通不好。